Mix gently by pipetting up and down or flicking the tube 4-5 times. Do not vortex. Place the mixture on ice for 30 minutes. Do not mix.

Heat shock at 42°C for 30 seconds. Do not mix.

Transfer tubes on ice for 2 minutes.

Add 950 µl of room temperature SOC media to tubes.

Place the tube at 37°C for 60 minutes. Shake vigorously (250 rpm) or rotate.

Warm selection plates to 37°C.

Spread 100 µl of the cells onto the plates with appropriate antibiotics. Use amp plates for positive control sample.

Incubate plates overnight at 37°C.

Qiagen MiniPrep

For overnight cell cultures 1~5mL of E. Coli in LB medium:

Transfer cell culture into centrifuge tubes

Centrifuge for 5 min. at 4,000 rpm

Discard the supernatant

Resuspend pelleted bacterial cells in 250 µl Buffer P1 and transfer to a microcentrifuge tube. Ensure that RNase A has been added to Buffer P1. No cell clumps should be visible after resuspension of the pellet.

Add 250 µl Buffer P2 and mix thoroughly by inverting the tube 4–6 times. Do not vortex. Continue inverting the tube until the solution becomes viscous + slightly clear blue. Do not allow lysis reaction to proceed for more than 5 min.

Add 350 µl Buffer N3 and mix immediately + thoroughly by 4-6x inverting the tube. The solution should become cloudy.